Identification, Validation and Classification of the Genus Phyllanthus in Nigeria Using ITS Genetic Marker and the Taxonomic Implication

Abstract

An extraction, purification, PCR amplification and sequencing of DNA from five species of Phyllanthus in Nigeria namely P. amarus Schum and Thonn, P. urinaria Linn., P. odontadenius Mull-Arg., P. niruroides Mull-Arg. and P. muellerianus (O. Ktze) Excel belonging to the family of Phyllanthaceae were carried out using nuclear ribosomal Internal Transcribed Spacer (ITS 4-5) genetic marker to identify unknown Phyllanthus species. The nuclear region revealed that the Phyllanthus species were able to be amplified optimally for sequencing. The results of the nucleotide sequences were further compared on Basic Local Alignment Sequence Tool (BLAST) on GenBank and BoldSystems for validation. Results revealed that the closely related species, P. niruroides Mull_Arg. and P. odontadenius Mull-Arg. had no DNA record to separate them on both GenBank and BoldSystems while P. amarus Schum and Thonn, P. muellerianus (O. Ktze) Excel and P. urinaria Linn. were clearly compatible with other works. The sequence data were analyzed and classified with tree-based analyses of Mr.Bayes 3.2.1. in order to reveal their phylogenetic relationship. Results of the nucleotide sequences and fragment analysis were published on BoldSystems for barcoding as non-coding marker translation matrix.